Background Increasing evidence links early-life exposure to psychosocial stress with adverse childhood respiratory outcomes. The influence of exposure timing has not been completely elucidated. Objective To examine the association between prenatal and postnatal maternal stress and wheeze in 417 children enrolled in a prospective birth cohort in Mexico City. Methods Maternal negative life event (NLE) scores were ascertained in the second or third trimester of pregnancy and at the 48-month postnatal visit. Children's respiratory outcomes, caregiver report of ever wheeze, and wheeze in the past 12 months were obtained from the International Study of Asthma and Allergies in Childhood survey administered at 48 months. Associations between prenatal and postnatal NLE scores and wheeze were analyzed using a modified Poisson regression approach adjusting for covariates. Results In separate models, higher maternal psychosocial stress during pregnancy (relative risk [RR], 1.12; 95% CI, 1.00-1.26) and postnatally (RR, 1.21; 95% CI, 1.08-1.35) were associated with increased risk of wheeze in the past 12 months with an evident exposure-response relationship. There was a significant interaction between postnatal stress and sex in relation to current wheeze. In a sex-stratified model, the association between postnatal stress and risk of wheeze in the past 12 months was stronger in girls (RR, 1.35; 95% CI, 1.13-1.61) than in boys (RR, 1.11; 95% CI, 0.97-1.27) (P for interaction =.04). Conclusion Prenatal and postnatal stress in mothers was associated with wheeze in preschool-aged children, and the effect of postnatal stress was stronger in girls. Understanding the temporal- and sex-specific effects of stress may better inform prevention strategies.